The co-effect of protective colloids and polylactic acid (PLA) on the polymorphic crystal forms and crystallinity of indomethacin (IMC) in IMC-loaded PLA microspheres was investigated with differential scanning calorimetry, infrared spectroscopy and x-ray diffractometry, to evaluate the polymorphic crystal forms and crystallinity of IMC encapsulated in PLA microspheres. The surfactant, sodium dodecyl sulphate (SDS), was also used as a dispersing agent. The results indicate that the polymorphism and crystallinity of IMC encapsulated in IMC-loaded PLA microspheres was dependent on the type of protective colloid and PLA used. The amorphous state and alpha-form of IMC were found in the IMC-loaded PLA microspheres prepared using polysaccharide (pectin or beta-cyclodextrin) as a protective colloid or SDS as a dispersing agent. However, the amorphous and methylene chloride solvate of IMC seemed to exist in the IMC-loaded PLA microspheres prepared with the proteins (gelatin or albumin), synthetic cellulose derivative (methyl cellulose or hydroxylpropyl methylcellulose) or the synthetic nonionic polymer (polyvinyl alcohol, polyvinyl pyrrolidone or biosoluble polymer) as a protective colloid. PLA was found to express a certain crystallinity in microspheres and not be affected by the protective colloids, but it played a more important role in influencing the crystallization of IMC during microencapsulation than the protective colloids. No interaction occurred in the physical mixture of IMC and PLA, nor in the IMC-loaded PLA microspheres.